CAJ | 학술논문

目的采用2-DE和质谱技术分析RA患者和创伤件关节炎患者关节滑膜组织中FLS蛋白质组学的变化,寻找RA关节滑膜特异性自身抗原.方法采用2-DE分离6例创伤性关节炎患者及3例RA患者关节滑膜组织中FLS总蛋白质,凝胶经银染显色和图像数据分析识别差异蛋白质点,并用MALDI-TOF-MS对筛选出的差异蛋白质点进行鉴定.结果 RA患者与创伤性关节炎患者中的FLS蛋白质通过2-DE检测,分别检出1 633、1 603个蛋白质点;对2组相互匹配的蛋白质点进行差异分析后,其中蛋白质差异量大于3倍的差异点共有92个,从中选取在RA患者FLS中表达上调的33个差异蛋白质点进行MALDI-TOF-MS 鉴定分析,并与IPI和Swiss-Prot数据库进行比对,共鉴定出包括肿瘤型丙酮酸激酶、α-烯醇化酶、二硫键异构酶A3 前体、谷胱甘肽转移酶、泛素结合酶E2K、血小板活化因子乙酰水解酶、二氢嘧啶酶样2、腺苷酸酶、转醛醇酶、乙酰基辅酶A异构酶、26S蛋白调节亚单位S6a、膜联蛋白A11、过氧化物还原酶1、人色氨酰tRNA合成酶、核纤层蛋白、肌间蛋白、核基质蛋白、肌动蛋白、T-复合物多肽1、葡萄糖调节蛋白75、αB-晶体蛋白、葡萄糖调节蛋白78、未知蛋白:如PSMFA和SELENBP1等30个与RA发病有关的蛋白.结论 RA患者与创伤性关节炎患者FLS间存在差异表达的蛋白质,这些差异蛋白有可能是构成RA关节滑膜的相关自身抗原,并可能在诱导RA自身免疫反应中发挥一定作用.
목적 채용2-DE화질보기술분석RA환자화창상건관절염환자관절활막조직중FLS단백질조학적변화,심조RA관절활막특이성자신항원.방법 채용2-DE분리6례창상성관절염환자급3례RA환자관절활막조직중FLS총단백질,응효경은염현색화도상수거분석식별차이단백질점,병용MALDI-TOF-MS대사선출적차이단백질점진행감정.결과 RA환자여창상성관절염환자중적FLS단백질통과2-DE검측,분별검출1 633、1 603개단백질점;대2조상호필배적단백질점진행차이분석후,기중단백질차이량대우3배적차이점공유92개,종중선취재RA환자FLS중표체상조적33개차이단백질점진행MALDI-TOF-MS 감정분석,병여IPI화Swiss-Prot수거고진행비대,공감정출포괄종류형병동산격매、α-희순화매、이류건이구매A3 전체、곡광감태전이매、범소결합매E2K、혈소판활화인자을선수해매、이경밀정매양2、선감산매、전철순매、을선기보매A이구매、26S단백조절아단위S6a、막련단백A11、과양화물환원매1、인색안선tRNA합성매、핵섬층단백、기간단백、핵기질단백、기동단백、T-복합물다태1、포도당조절단백75、αB-정체단백、포도당조절단백78、미지단백:여PSMFA화SELENBP1등30개여RA발병유관적단백.결론 RA환자여창상성관절염환자FLS간존재차이표체적단백질,저사차이단백유가능시구성RA관절활막적상관자신항원,병가능재유도RA자신면역반응중발휘일정작용.
Objective To identify synovium-associated proteins of by analyzing proteome of FLS between RA patients and traumatic arthritis. Methods The total protein of FLS from 3 patients of RA and 6 normal controls were separated by 2-DE. The differentially expressed proteins were identified by MALDI-TOF-MS. Results The 2-DE image results showed that there were many proteins differentially expressed in FLS from RA and control. A total of 33 differential proteins that were up-regulated in RA as compared with controls were selected and 30 proteins including pyruvate kinase isozymes M1/M2, α-enolase, protein disulfide-isomerase A3 precursor, glutathione transferase, peroxiredoxin 1, ubiquitin-conjugating enzyme E2K, platelet-activating factor acetylhydrolase IB, dihydropyrimidinase-related protein 2, adenosine kinase, transaldolase, δ-δ-dienoyl-CoAisomerase, 26S protease regulatory subunit 6A, annexin All, tryptophanyl-tRNA synthetase isoform b, lamin-A/C, myomesin-1 isoform b, ruvB-like 1, actin-cytoplasmic 1, T-complex protein 1, stress-70 protein, mitochondrial precursor, α-crystallin B chain, 78 000 glucose-regulated protein, PSME4, SELENBP1 and so on. Conclusion The differentially expressed proteins are present in FLS from RA and control, which may be the synovium biomakers and involved in pathogenesis of RA.